Egypt. Acad. J. biolog. Sci., 3 (1): 213 - 22 0 (2010) A. Entomology
Email: egyptianacademic@yahoo.com ISSN: 1687–8809
Received: 29/6 /2010 www.eajbs.eg.net
Effect of different feeding diets on the haemolymph of the newly emerged
honeybee workers Apis mellifera L.
Sawsan S. El Mohandes; Emad A. Nafea and Asmaa M. Fawzy
Department of Apiculture, Plant Protection Research Institute, Agriculture
Research Center, Dokki, Giza, Egypt.
ABSTRACT
Five diets were compared for measuring their effects on the differential
haemocytes types of the newly emerged worker bees, and also on their haemolymph
proteins, lipids and glucose content. As blood haemocytes cell play a role in
defending honeybees against parasites and pathogens. Five groups of newly emerged
worker bees were fed with one of the following diets in patty form with sugar: faba
bean (Vicia faba) pollen, maize (Zea mays) pollen, date palm (Phoenix dactylifera)
pollen, Egyptian clover (Trifolium alexandrinum) pollen and soya bean (Glycine max)
flour (as pollen supplement). Differential haemocyte counts (DHC) were evaluated in
smears of their haemolymph. The types of the haemocytes (prohaemocytes,
plasmatocytes, oenocytoids, granulocytes, coagulocytes and binucleated cells) were
recorded. The most abundant type was plasmtoycte cells (over 90%) followed by
granulocyte cells and coagulocyte cells. These blood cells perform phagocytosis and
encapsulation of foreign bodies in the honeybee body cavity. Feeding honeybees with
these different diets caused significant differences between the haemocyte cells. The
highest number of plasmatocyte cells recorded in bees which fed on maize pollen,
while the lowest was found in bees fed on bean pollen. The highest protein content
was found in the haemolymph of bees fed on date palm and the lowest was found in
bees fed on bean pollen and the difference was non-significant. The difference in
lipids content was non-significant in worker bees fed on these different diets. There
was a significant difference in glucose content between bees fed on maize pollen and
date palm pollen.
Keywords: honeybee, haemolymph, beedieds
INTRODUCTION
Insect’s haemolymph plays a very important role in transport and storage of
nutrients and is crucial for the recognition and defense against micro-organism
(Bogaerts et al., 2009). Honeybees possess an open circulatory system and numerous
haemocytes are contained in its haemolymph. Prohaemocytes, plasmatocytes, granular
cells, cystocytes, sphaerula cells and oenocytoids are the cells that comprise the bee
haemocyte population (Gupta, 1991). They are variable in morphology and functions.
They are responsible for the defense reactions against foreign agents that penetrate the
haemocoel (Tepass et al., 1994; Falleiros and Gregorio, 1995; Gliński and Jarosz,
1995; and Inoue et al., 2001). They perform phagocytosis, encapsulation of foreign
bodies in the insect body cavity, coagulation to prevent loss of blood, nodule
formation, transport of food material, (Patton, 1983). The haemocytes can engulf and
destroy smaller foreign objects such as bacteria or fungal spores, but larger parasites,
Sawsan S. El Mohandes et al.214
bacterial clumps or fungal hyphae, are encapsulated by several haemocytes and then
removed from circulation (Gliński and Jarosz, 2001).
Honey bees require proteins, carbohydrates, lipids, vitamins, minerals,
and water in their diets. As protein plays a major role in the life of honey bees
(Amdam and Omholt, 2002), colonies that have no access to pollen have reduced
capacity to rear brood, quickly decline in population, and may eventually die. Protein
deficiency also affects the ability of honey bees to resist diseases (Matilla and Otis,
2006) and causes a major disturbance in the structure and functioning of the
haemolymph cellular system (Rogala and Szymas, 2004). Lipids are also important
for the functioning of cellular membranes. Carbohydrates form a large part of the diet
of the colony and are required by both the larva and adult for normal growth and
development. Carbohydrates in the bee’s diet are used mainly to generate energy for
muscular activity, body heat, and vital functions of certain organs and glands, such as
wax production (Standifer et al., 1977)
The objective of this study was to evaluate the influence of different feeding
diets (pollen grains and pollen substitute) into the differential haemocyte count (DHC)
in the haemolymph of the newly emerged worker honeybees, once that the success of
the immune response in insects depends on the number and the types of haemocytes
(Russo et al., 2001). Also the different diets effects on proteins, lipids and glucose
content in their haemolymph.
MATERIALS AND METHODES
This study was carried out in the Apiary of Beekeeping Research Department,
Plant Protection Research Institute during the late summer of 2009 till summer 2010.
There were five experimental groups each group contained three colonies of equal
strength. Each group fed with one of the following pollen grain cakes by mixing 50gm
of each pollen with 100gm powdered sugar. Distilled water was added to obtain a
patty. The pollen grains used in this experiment were collected by honeybees from
four plants in different regions of Giza Governorate. These pollen grains were faba
bean (Vicia faba), maize (Zea mays), date palm (Elaies guineensis), Egyptian clover
(Trifolium alexandrinum) and soya bean (Glycine max) flour. These pollen grains
were collected from pollen traps and stored in a deepfreeze at about -4ºC. The diet
cakes were placed directly over the center of the brood combs and the top of the cake
was covered with waxed paper to prevent moisture loss. The experimental colonies
were subjected to these different feeding treatments every week begin from late
summer of 2009 till summer 2010. Investigated bees were collected by placing frames
with emerging brood from each colony fed with these feeding treatment in incubators
set to the temperature of 33°C and the relative humidity of 65% till the worker
honeybee emerged. A drop of the haemolymph of the newly emerged worker was
taken over a clean microscope slide and a smear was made. Smears were stained with
Giemsa stain and differential haemocyte counts (DHC) were evaluated. One hundred
cells were counted per slide under oil immersion and phase contrast using light Leitz
microscope. The types of the haemocytes (prohaemocyte, plasmatocytes,
granulocytes, coagulocyte and oenocytoid) were recorded using the key of Gupta
(1979). About 30 smears of haemolymph for each diet were examined. Chemical
analysis was done on the haemolymph of pooled sample collected from newly
emerged worker bees and kept in deep freezer till analysis. Total protein content was
determined by Biuret reagent according to Gornal, et al., (1949) method. Total lipids
Effect of different feeding diets on the haemolymph of the newly emerged honeybee 215
were determined using Knight et al., (1972) method while total carbohydrate was
determined using Trinder (1969) method. The statisitical analysis including one way
ANOVA at Ρ < 0·05 level were done using SPSS 10 software.
RESULTS AND DISCUSSION
Six haemocyte types were observed in the haemolymph of the newly emerged
worker bees feeding on different types of diet and were represented in Table (1) and
Figure (1). They were classified as prohaemocytes, plasmatocytes, granulocytes,
coagulocytes, oenocytoid and binucleated cells.
Table 1: Effect of different feeding pollen diets on different haemocytes count of newly emerged
worker of honeybee
(PR, prohamocyte; PL, plasmatocyte; GR, granulocyte; CO, coagulocyte, OE, oenocytoid,
Bin, binucleated cells)
* indicates significant differences (P < 0.05) between the mean values of the haemocytes
** SE= standard error
Fig. 1: Haemocyte types in the haemolymph of the newly emerged worker honeybees
Diet Percentage of haemocyte types (mean ± SE)**
PR PL GR CO OE Bin
Faba Bean
pollen
2.13
±1.08
90.29*
±3.09
5.38*
±1.30
1.50*
±1.01
0.38*
±0.15
0.33*
±0.21
Maize
pollen
0.0 97.05*
±0.83
1.70
±0.24
0.97*
±0.53
0.08*
±0.003
0.20*
±0.15
Date palm
pollen
0.0 93.95
±0.97
3.39
±1.30
2.33
±0.19
0.0 1.33*
±0.33
Clover pollen 1.20
±0.61
90.27*
±1.52
2.90
±0.10
5.07*
±2.13
0.33*
±0.12
0.23*
±0.003
Soya bean
flour
0.0 93.01
±0.66
2.72
±0.73
4.07
±0.41
0.13
±0.01
0.08*
±0.01
PR= Prohaemocyte PL= Round Plasmatocyte PL= Oval Plasmatocyte
PL= Irregular Plasmatocyte GR= Granulocyte CO= Coagulocyte
OE= Oenocytoid Bin= Binucleated cell
Sawsan S. El Mohandes et al.216
Prohaemocytes (PR) are mostly appear rounded in shape but some are oval or
spindle – shaped, and small in size. The nucleus fills almost the entire cell and is
mostly centrally located. The cytoplasm is restricted to a thin layer around the nucleus
showing pronounced basophilia. It contains small granules, droplets or vacuoles.
Plasmatocytes (PL) are larger than prohaemocytes with variable sizes, round, oval or
spindle in shape and sometimes irregular with projections. The nucleus is round or
elongated in shape and is generally centrally located with a distinct nucleolus. The
cytoplasm is abundant basophilic and may exhibit small vacuoles. Granulocytes (GR)
are rounded to ovoid cells and the nucleus is relatively small, rounded or elongated
and is centrally located. The cytoplasm is characteristically granular. Coagulocytes
(CO) are spherical or oval cells. The nucleus is relatively small, rounded or elongated
and is centrally located. The cytoplasm is characteristically or irregular polygonal.
Oenoctoids (OE): The OEs are rounded or oval in shape, and mostly large cells.
Compared to cell size, the nucleus is small and is eccentrically located. It exhibits a
moderate acidophilia, and reveals a homogenous cytoplasm containing fine and weak
acidopholic granulation. Binucleated cells (Bin): It seemed to represent incomplete
mitosis of cell division according to Arnold and Hinks (1976).
The predominant type of the haemocytes of bees fed with the five tested diets
types was the plasmatocyte cells followed by the granulocyte, and coagulocyte.
These results are in agreement with those of previous studies which mentioned that
plasmatocytes were the most numerous cells in the haemolymph of young honeybee
from free-flying colonies (Jędruszuk, 1998a, b and c and Szymas and Jędruszuk,
2003). Other haemocyte cells like the prohaemocyte, oenocytoid and binnucleated
cells were less abundant or not present in all the feeding type. The high percentage in
plasmatocytes cells was found in bees fed on maize pollen (97.05), while the lower
percentage was found in bees fed on clover pollen (90.27). There were significant
differences between the effect of maize pollen and both of faba bean and clover
pollen. Vice versa the highest percentage of granular haemocyte was found in worker
bees fed on faba bean pollen (5.38) and the lowest percentage was found in bees fed
on maize pollen (1.70) with significant difference between them. The high percentage
of coagulocyte cells were present in bees fed on clover pollen while the low
percentage was found in bees fed on maize pollen. There were significant differences
between clover pollen and both of bean and maize pollen. Prohaemocyte cells present
only in bees fed on faba bean and clover pollen and were absent in worker bees fed on
maize pollen, date palm pollen and soya bean flour. The high percentage of
prohaemocyte cells was found in bees fed on faba bean pollen. Oenocytoid cells were
observed in the haemocyte of bees except these fed on date palm pollen where it was
absent. The higher percentage of oenocytoid cells was found in bees fed on faba bean
pollen (0.38%) and a lower percentage was found in bees fed on maize pollen and the
difference between them was significant. The highest percentage of binucleated cells
was found in date palm pollen and the lower percentage was found in bees fed on
soya bean flour. There was a significant difference between the date palm pollen and
all the other different diets.
The cellular immune reactions have been shown to be accompanied by
changes in both of the number of circulating haemocytes and in the relative
proportions of different haemocyte types in the blood (Hink, 1970). From the previous
results it was noticed that the plasmatocyte are the most abundant cells followed by
granulocyte and coagulocyte cells. These results agreed with those of Szymaś and
Jedruszuk, (2003). They mentioned that there was a significantly higher percentage in
Effect of different feeding diets on the haemolymph of the newly emerged honeybee 217
plasmatocytes and a significant decrease of granular haemocytes in the haemolymph
of bees fed with pollen substitute, compared with the haemocyte system of bees fed
with pollen. As phagocytosis and encapsulation are two common types of defense
reactions in honeybee against invading pathogens. Encapsulation began when
granular cells attached to the foreign target and this was followed by attachment of
multiple layers of plasmatocytes to form the outer layer of the capsule (Louis and
Michael, 1996; Gliński and Jarosz, 2001 and Glinski and Buczek, 2003). The
physiological mechanisms effect of phagocytosis, encapsulation, and other related
defense mechanisms was due to the availability of circulatory immune cells
particularly plasmatocytes and granulocytes (Sanjayan, et al, 1996). Bee haemocytes
may directly kill bacteria, fungal spores and other small foreign molecules in
phagocytic process (Gotz, 1986).
The influence of the different diets on the haemolymph proteins, lipids and
glucose of the newly emerged worker honeybee was studied and was presented in
Table (2). It was noticed that the total protein content in the haemolymph of the newly
emerged worker fed on the five diets were varied in their values but with no
significant between them.
Date palm pollen was the highest (10.48%) in haemolymph protein content
followed by soya bean flour (9.77%), while faba bean pollen, maize pollen and clover
pollen were similar in their haemolymph protein and were the lowest. The natural
protein is an important factor in the functioning of the haemolymph cellular system of
bees (Szymaś and Jedruszuk, 2003).
Concerning to the total lipid contents in the newly emerged workers, results in
table (2) showed no significant differences between the five tested diets. It ranged
between 48.36% in date palm pollen and 50.0% in soya bean flour.
Table 2: Protein, lipids and glucose content in the haemolymph of the newly emerged worker bees fed
on different diets (mean value ±S.E.).
± S.E. = standard error values
* indicates significant differences (P < 0.05) between the mean values
Lipids are used by larvae and young adult bees as sources of energy and for
the synthesis of reserve fat and glycogen. For that young adult bees require and utilize
some of the lipids in the pollen they consume. Manning (2001) mentioned that pollens
with high lipid concentrations and dominated by linoleic, linolenic, myristic and
dodecanoic acids probably play a significant role in inhibiting the growth of the sporeforming
bacteria, Paenibacillus larvae (American foulbrood), Melissococcus pluton
(Europeanlarvae larvae foulbrood) and other microbes that inhabit the brood combs of
bee hive
Table (2) also represents the haemolymph glucose content of the newly
emerged worker honeybees. The highest value was found in workers fed on maize
pollen (7.38%) while the lowest content was recorded in worker bees fed on date palm
pollen (6.86%) with a significant difference between them. Other treatments were
Type of Diets Protein mg/ml Lipid mg/ml Glucose mg/ml
Faba bean pollen 8.28 ±0.31 49.29 ±0.36 7.06 ±0.19
Maize pollen 8.74 ±0.30 49.84 ±0.16 7.38*±0.01
Date palm pollen 10.48 ±1.17 48.36 ±1.4 6.86*±0.23
Clover pollen 8.97 ±0.58 49.76 ±0.14 7.22 ±0.02
soya bean flour 9.77 ±0.80 50.0 ±0.00 7.23 ±0.01
Sawsan S. El Mohandes et al.218
arranged in between with no significant differences between them and the glucose
content on these fed on maize pollen. The main energy source for social insects was
carbohydrates and it provides adult honey bees with the energy to fly and exist. The
glucose was taken as example to carbohydrate.
The forementioned results proved the important role of the nutritional value of
feeding diets on the haemocyte types, and its proteins, lipids and carbohydrates
content. The most abundant haemocyte cells were the plasmtocytes followed by the
granular cells. Haemocytes play an essential role in defending insects against invading
parasites and pathogens. The effective physiological mechanisms of phagocytosis,
encapsulation, and other related defense mechanisms were primarily due to the
availability of circulatory immune cells particularly plasmatocytes and granulocytes.
Feeding honeybee with these different diets may help enhancing the immunity of the
bees due to their positive effects on the types and percentage of the haemocytes.
Accordingly, it is premise feeding honeybee with pollen grain or their supplement to
increase their defense reaction against different diseases.
REFERENCES
Amdam, G.V. and Omholt, S.W. (2002). The regulatory anatomy of honey bee
lifespan. Journal of Theoretical Biology, 216: 209-228.
Arnold, J.W. and Hinks, C.F. (1976). Haemopoiesis in Lepidoptera. I. The
multiplication of circulating haemocytes. Can. J. Zool., 54: 1003-1012.
Bogaerts, A.; Baggerman, G.; Vierstraete, E.; Schoofs, L. and Verleyen, P. (2009).
The hemolymph protein of the honeybee: Gel-based or gel-free? Proteomics,
9: 3201- 3208.
Falleiros, A. M. F. and Gregorio, E. A. (1995). Hemocitos fugocitarios em larvas de
Diatraea saccheralis (Fabricius) (Lepidoptera, Pyralidae). Revista Brasileira de
Zoologia, 12 (4): 751–758.
Gornal, A.G; Bardawil, C.J. and David, M.M. (1949). Determination of serum protein
by means of the biuret reaction J. Biol. Chem., 177: 751.
Gliński, Z. and Buczek, K. (2003). Response of the Aoidea to fungal infections.
Apiacta, 38: 183-189.
Gliński, Z. and Jarosz, J. (1995). Cellular and humoral defences in honey bees. Bee
world, 67:195-205
Gliński, Z. and Jarosz, J. (2001). Infection and immunity in the honey bee, Apis
mellifrea L., Apiacta, 36 (1): 12-24.
Götz, P. (1986). Encapsulation in Arthropods. In Brehèlin, M; Boemare N. (eds)
Immunity in Invertebrates. Springer Verlag, Berlin, Heidelberg, pp. 153-170
Gupta, Α. Ρ. (1979). Haemocyte types, their structure, species interrelationship and
taxonomic significance; in Insect haemocytes (Cambridge: Cambridge
University Press) pp. 85-127.
Gupta A.P., Ed. (1991). Immunology of Insects and other Arthropods. CRC Press,
Boca Raton, FL.
Hink, W.F. (1970) Immunity in insects. Transplantation Proceedings 2: 233 - 235
Inoue, N.; Hanada, K.; Natoshi, T.; Igarashi, I.; Nagasawa, H.; Mikami, T. and
Eujisaki, K. (2001). Characterization of phagocytic hemocytes in
Ornithodoros moubata (Acari: Ixodidae). J. Medical Entomology, 35 (4): 514-
519.
Effect of different feeding diets on the haemolymph of the newly emerged honeybee 219
Jędruszuk A. (1998a). Age-dependence of metabolic activity of haemocytes in
haemolymph of adult generation, Pszcz. Zesz. Nauk. XLII 2: 41- 42.
Jędruszuk A. (1998b). Total haemocyte counts in haemolymph of adult worker honey
bees (Apis mellifera L.) from spring-summer and autumnwinter generations,
Pszcz. Zesz. Nauk.XLII 2: 43- 44.
Jędruszuk A. (1998c). Wstępna analiza zmian obrazu hemolimfy pod wpływem wieku
u pszczół robotnic A. mellifera, XXXV Nauk. Konf. Pszczel. Puławy 11-12
marca 1998, pp. 32-33.
Knights, J.A.; Anders, S. and Rawle, J.M. (1972). Chemical basis of the sulfophospho-vanillin
reaction for estimating total serum lipids. Clin. Chem.
18:199-202.
Louis, L. P. and Michael, R. S. (1996). Granular cells are required for
encapsulation of foreign targets by insect haemocytes. J. Cell Science 109:
2053-2060.
Manning, R (2001). Fatty acids in pollen: a review of their importance for honey bees.
Bee World, 82 (2): 60-75.
Matilla, H. R. and Otis, G. W. (2006). Effects of pollen availability and Nosema
infection during the spring on division of labour and survival of worker honey
bees (Hymenoptera: Apidae). Environmental Entomology 35: 708-717.
Patton, R. L., (1983). Introductory insect physiology. W. B. Saunders Co.
Philadelphia. 47, 65 pp.
Rogala, R. and Szymas, B. (2004). Nutritional value for bees of pollen substitute
enriched with synthetic amino acids Part II. Biological methods. J. Apicultural
Science, 48 (1): 1-8.
Russo, J.; Brehelin, M. and Carton, Y. (2001). Hemocyte changes in resistant and
susceptible strains of D. melanogaster caused by virulent and a virulent strains
of the parasitic wasp Leptopilina bouladi. Journal of Insect Physiology 47:
167-172.
Sanjayan, K.P.; Ravikumar, T. and Albert, S. (1996). Changes in the haemocyte
profile of Spilostethus hospes (Fab) (Heteroptera: Lygaeidae) in relation to
eclosion, sex and mating. J. Biosci., 21(6): 781-788.
Standifer, L. N.; Moeller, F. E.; Kauffeld, N. M.; Herbert, E. W.; Jr., and Shimanuki,
H. (1977). Supplemental feeding of honey bee colonies. United States
Department of Agriculture, Agriculture Information Bulletin 413: 8 pages,
illus.
Szymas, B. and Jêdruszuk, A. (2003). The influence of different diets on haemocytes
of adult worker honey bees, Apis mellifera. Apidologie 34: 97-102
Tepass, U.; Fessler, L. I.; Aziz, A. and Hestensein, V. (1994): Embryonic origin of
hemocytes and their relationship to cell death in Drosophila Development.
120: 1829-1837.
Trinder, P. (1969): Enzematic determination of sugar in serum and plasma. Amm.
Clin. Biochem., 6: 24.
Sawsan S. El Mohandes et al.220
ARABIC SUMMARY
‫الخروج‬ ‫حذيثة‬ ‫النحل‬ ‫شغاالت‬ ‫دم‬ ً‫عل‬ ‫المختلفة‬ ‫التغذية‬ ‫تاثير‬Apis mellifera L.
‫المهنذس‬ ‫سعيذ‬ ‫سىسن‬-‫نافع‬ ‫عماد‬-‫فىزي‬ ‫محمذ‬ ‫أسماء‬
‫البح‬ ‫مزكش‬ ،‫النباحاث‬ ‫وقايت‬ ‫بحىد‬ ‫معهد‬ ،‫النحل‬ ‫بحىد‬ ‫قسم‬‫مصز‬ ، ‫الشراعت‬ ‫وسارة‬ ، ‫الشراعيت‬ ‫ىد‬
‫صيف‬ ‫اواخز‬ ً‫ف‬ ً‫بالدق‬ ‫النحل‬ ‫قسم‬ ‫منحل‬ ً‫ف‬ ‫الدراست‬ ‫هذة‬ ‫أجزيج‬9002‫صيف‬ ‫نهايت‬ ً‫حخ‬9000.‫وقدد‬
‫دذل‬‫د‬‫وك‬ ‫دزوس‬‫د‬‫الم‬ ‫دديذ‬‫د‬‫ح‬ ‫النحدل‬ ً‫د‬‫د‬‫ف‬ ‫ددم‬‫د‬‫ال‬ ‫يدا‬ ‫خ‬ ‫دىال‬‫د‬‫ان‬ ً‫د‬‫د‬ ‫ع‬ ‫دا‬‫د‬‫حاريزه‬ ‫ورراسدت‬ ‫دت‬‫د‬‫الخاذي‬ ‫دة‬‫د‬‫م‬ ‫اندىال‬ ‫دت‬‫د‬‫س‬ ‫خ‬ ‫دخمدام‬‫د‬‫أس‬ ‫حدم‬
‫وا‬ ً‫والدهن‬ ً‫البزوحين‬ ‫حخىي‬ ‫ال‬ ً ‫ع‬ ‫حاريزها‬‫رمائها‬ ً‫ف‬ ‫لكزبىهيدراث‬.‫الخةزبدت‬ ‫هدذة‬ ً‫فد‬ ‫اسدخمدمج‬01‫نحدل‬ ‫يدت‬ ‫خ‬
ً‫د‬‫د‬‫ال‬ ‫ج‬ ‫د‬‫د‬‫قس‬1‫دت‬‫د‬‫ىع‬ ‫مة‬ ‫دل‬‫د‬‫ك‬ ‫داث‬‫د‬‫ىع‬ ‫مة‬3‫دا‬‫د‬‫ي‬ ‫خ‬‫دى‬‫د‬‫ف‬ ‫دا‬‫د‬ ‫ل‬ ‫دى‬‫د‬‫حب‬ ً‫د‬‫د‬ ‫ع‬ ‫دذيخها‬‫د‬‫حا‬ ‫دم‬‫د‬‫ح‬ ً‫د‬‫د‬‫الول‬ ‫دت‬‫د‬‫ىع‬ ‫ة‬ ‫ال‬(10‫دم‬‫د‬‫ج‬)
‫بدىررة‬ ‫دكز‬‫د‬‫س‬ ‫مدس‬ ‫ىطدت‬ ‫مم‬(000‫جدم‬)‫دا‬‫د‬ ‫ال‬ ‫مدة‬ ‫دل‬‫د‬‫ي‬ ‫وق‬.‫دن‬‫د‬‫ب‬ ‫رة‬ ‫دا‬ ‫ل‬ ‫دى‬‫د‬‫حب‬ ً‫د‬ ‫ع‬ ‫حادذيخها‬ ‫دم‬‫د‬‫ح‬ ‫الزانيدت‬ ‫ىعدت‬ ‫ة‬ ‫ال‬
‫السداب‬ ‫النسب‬‫ت‬.‫ت‬ ‫السداب‬ ‫النسدب‬ ‫بدن‬ ‫النميدل‬ ‫دس‬ ‫ط‬ ‫دا‬ ‫ل‬ ‫حبدى‬ ً‫د‬ ‫ع‬ ‫حادذيخها‬ ‫حدم‬ ‫الزالزدت‬ ‫ىعدت‬ ‫ة‬ ‫وال‬.‫ىعدت‬ ‫ة‬ ‫وال‬
‫ت‬ ‫الساب‬ ‫النسب‬ ‫بن‬ ‫البزسيم‬ ‫ا‬ ‫ل‬ ‫حبى‬ ً ‫ع‬ ‫حاذيخها‬ ‫حم‬ ‫الزابعت‬.‫رقيد‬ ً‫د‬ ‫ع‬ ‫حادذيخها‬ ‫حم‬ ‫د‬ ‫ف‬ ‫المامست‬ ‫ىعت‬ ‫ة‬ ‫ال‬ ‫اما‬
‫ت‬ ‫الساب‬ ‫النسبت‬ ‫بن‬ ‫الصىيا‬ ‫فى‬.‫النحل‬ ‫رم‬ ‫يا‬ ‫خ‬ ‫انىال‬ ‫فحص‬ ‫عند‬‫المارس‬:‫وجددث‬ ‫ىعدت‬ ‫مة‬ ‫لكل‬ ‫حديزا‬6‫أندىال‬
ً‫وهدد‬ ‫الدددم‬ ‫يددا‬ ‫خ‬ ‫مددة‬‫اث‬ ‫يددت‬ ‫خ‬ ،‫كىاجيىلىسدديج‬ ،‫جزانيىلىسدديج‬ ،‫اوينىسدديخىيد‬ ،‫سماحىسدديج‬ ‫ب‬ ،‫ىسدديج‬ ‫بزوهي‬
‫نىاحية‬. )‫مدة‬ ‫اسيدد‬ ‫سماحىسدايج‬ ‫الب‬ ‫يدا‬ ‫خ‬ ً‫فد‬ ‫موىيدت‬ ‫نسدبت‬ ً ‫أع‬ ‫وجدث‬ ‫وقد‬20%‫جزانيىلىسديج‬ ‫يدا‬ ‫خ‬ ‫يهدا‬ ‫ح‬
‫الم‬ ‫دا‬‫د‬‫أم‬ ‫ديج‬‫د‬‫كىاجيىلىس‬ ‫دا‬‫د‬‫ي‬ ‫وخ‬‫دزي‬‫د‬‫الخ‬ ‫دت‬‫د‬‫الخاذي‬ ‫دىال‬‫د‬‫ان‬ ً‫د‬‫د‬‫حتهزف‬ ‫دم‬‫د‬‫ل‬ ‫أو‬ ‫دت‬‫د‬ ‫ي‬ ‫ق‬ ‫دبخ‬‫د‬‫نس‬ ‫دج‬‫د‬‫كان‬ ‫دد‬‫د‬ ‫ف‬ ‫دزي‬‫د‬‫الخ‬ ‫دا‬‫د‬‫ي‬.‫دد‬‫د‬‫وق‬
‫مخ‬ ‫ال‬ ‫الخاذيت‬ ‫بهذة‬ ‫حاذيخها‬ ‫عند‬ ‫الدم‬ ‫يا‬ ‫خ‬ ً‫ف‬ ‫معنىي‬ ‫فزوق‬ ‫وجدث‬‫سماحىسديج‬ ‫الب‬ ‫يدا‬ ‫خ‬ ً‫فد‬ ‫نسبت‬ ً ‫اع‬ ‫وكانج‬
(20.01)%‫حبدى‬ ً‫د‬ ‫ع‬ ‫ىدذي‬ ‫الدذي‬ ‫النحدل‬ ً‫ف‬ ‫وجدث‬ ‫موىيت‬ ‫نسبت‬ ‫وأقل‬ ‫الذرة‬ ‫ا‬ ‫ل‬ ‫حبى‬ ً ‫ع‬ ‫النحل‬ ‫حاذيت‬ ‫عند‬
‫البزسيم‬ ‫ا‬ ‫ل‬(20.90.) %‫الةزانيىلىسيج‬ ‫يا‬ ‫خ‬ ً‫ف‬ ‫نسبت‬ ً ‫أع‬ ‫وجدث‬ ‫وقد‬(1.35) %‫ىدذي‬ ‫الدذي‬ ‫النحدل‬ ً‫فد‬
‫د‬ ‫أع‬ ‫أمدا‬ ‫دى‬ ‫ال‬ ‫ا‬ ‫ل‬ ‫حبى‬ ً ‫ع‬‫دا‬ ‫ل‬ ‫حبدى‬ ً‫د‬ ‫ع‬ ‫ىدذي‬ ‫الدذي‬ ‫النحدل‬ ً‫فد‬ ‫كىاجيىلىسديج‬ ‫يدا‬ ‫خ‬ ً‫فد‬ ‫موىيدت‬ ‫نسدبت‬ ً
‫البزسيم‬(1.00.) %‫بحبدى‬ ‫الخاذيدت‬ ‫ان‬ ‫نةدد‬ ‫العسدل‬ ‫نحدل‬ ‫حندزة‬ ً‫فد‬ ‫الدفاعيدت‬ ‫يدا‬ ‫الم‬ ً‫هد‬ ‫يا‬ ‫الم‬ ‫هذة‬ ‫أن‬ ‫وحيذ‬
‫العسل‬ ‫نحل‬ ‫حنزة‬ ‫مناعت‬ ‫مة‬ ‫حشيد‬ ‫قد‬ ‫ت‬ ‫مخ‬ ‫ال‬ ‫ا‬ ‫ال‬.
‫السا‬ ‫النىال‬ ‫بهذة‬ ‫الخاذيت‬ ‫حاريز‬ ‫رراست‬ ‫وعند‬‫حديزا‬ ‫المارس‬ ‫النحل‬ ً‫ف‬ ً‫والدهن‬ ً‫البزوحين‬ ‫حخىي‬ ‫ال‬ ً ‫ع‬ ‫ت‬ ‫ب‬
‫سدت‬ ‫الم‬ ‫الندىال‬ ‫بهدذة‬ ‫الخاذيدت‬ ‫عند‬ ‫معنىي‬ ‫ىيز‬ ‫زق‬ ‫ال‬ ‫ان‬ ‫وجد‬.‫عندد‬ ‫معندىي‬ ‫فدزق‬ ‫وجدد‬ ‫ىكدىس‬ ‫الة‬ ‫حالدت‬ ً‫فد‬ ‫دا‬ ‫بين‬
‫الذرة‬ ‫ا‬ ‫ل‬ ‫حبى‬ ً‫ف‬ ً ‫أع‬ ‫كان‬ ‫وقد‬ ‫النميل‬ ‫س‬ ‫وط‬ ‫الذرة‬ ‫ا‬ ‫ل‬ ‫حبى‬ ‫مة‬ ‫بكل‬ ‫الخاذيت‬(0.35.)%
‫الدراسد‬ ‫هدذة‬ ‫ومة‬ً‫فد‬ ‫الدفاعيدت‬ ‫يدا‬ ‫الم‬ ‫مدة‬ ‫يشيدد‬ ‫دا‬ ‫ال‬ ‫حبدى‬ ‫بددائل‬ ‫أو‬ ‫دا‬ ‫ال‬ ‫بحبدى‬ ‫الخاذيدت‬ ‫أن‬ ‫نةدد‬
‫لألمزاض‬ ‫اومخها‬ ‫م‬ ‫مة‬ ‫يشيد‬ ً‫وبالخال‬ ‫العسل‬ ‫نحل‬ ‫حنزة‬.